Picturing the Membrane‐assisted Choreography of Cytochrome P450 with Lipid Nanodiscs

Abstract

Cytochrome P450, a family of monooxygenase enzymes, is organized as a catalytic metabolon, and requires enzymatic partners as well as environmental factors that tune its complex dynamic activity. P450 and its reducing counterparts are membrane-bound proteins which are believed to dynamically interact to form functional complexes. Increasing experimental evidence signifies the role (s) of protein-lipid interactions in P450's catalytic function and efficiency. The challenges posed by the membrane have severely limited high-resolution understanding of the molecular interfaces of these interactions. Nevertheless, recent NMR studies have provided piercing insights into the dynamic structural interactions that enable the function of P450. In this review, we will discuss different biomimetic approaches relevant to unveil molecular interplays at the membrane, focusing on our recent work on lipid-nanodiscs. We also highlight the need to expand the use of nanodiscs, and the power of a combination of cutting-edge solution and solid-state NMR techniques, to study the dynamic structures of P450 as well as other membrane-proteins.

The Role of the FMN-Domain of Human Cytochrome P450 Oxidoreductase in Its Promiscuous Interactions With Structurally Diverse Redox Partners

“…Moreover, the role of lipids and membrane anchoring of CPR and CYP have been recently confirmed to be of importance in the effectiveness of CPR's ET (Barnaba et al, 2017;Campelo et al, 2017;Denisov and Sligar, 2017; Barnaba and Ramamoorthy, 2018).…”

Influence of Transmembrane Helix Mutations on Cytochrome P450-Membrane Interactions and Function

“…A number of functional studies have demonstrated the catalytic competency of Nanodisc-incorporated CYPs (16,(43)(44)(45). The structural properties of CYPs incorporated in Nanodiscs have previously been investigated by a variety of experimental modalities, including small-angle x-ray scattering, solution and solid-state NMR (ssNMR), and atomic force microscopy (46)(47)(48). These studies all indicate that the N-terminal membrane anchor is embedded in the lipid bilayer of the Nanodisc.…”

Functional Assembly of Caenorhabditis elegans Cytochrome b-2 (Cecytb-2) into Phospholipid Bilayer Nanodisc with Enhanced Iron Reductase Activity

“…Nanodisc (ND) is a newly introduced approach to study membrane proteins which are reconstituted into a phospholipid bilayer encircled by two molecules of a membrane scaffold protein (MSP) as a belt. NDs could overcome all undesirable effects of detergent by providing native-like environments for the membrane proteins [14,15] including cytochromes P450 and b 5 [16][17][18][19][20]. Self-assembly of the membrane proteins in nanodiscs enables the usage of various biochemical and biophysical techniques for studying the structure and functions of the membrane proteins [21].…”